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Chapter 27: Amino Acids, Peptides, and Proteins.
monomer unit: -amino acids
H NH2
R
R = sidechain
CO2H
- Amino Acid
Biopolymer: the monomeric amino acids are linked through an
amide bond (the carboxylic acids of one AA with the -amino
group of a second)
R1
H3N
+
CO2
R2
H3N
R1
- H2O
H3N
CO2
O
N-terminus
O
H
N
R1
R2
N
H
O
H
N
O
H
N
R3
R4
N
H
O
H
N
O
R5
R6
N
H
CO2
O
H
N
O
C-terminus
R2
R7
N
H
Peptide or protein (polypeptide)
peptide (< 50 amino acids)
protein (> 50 amino acids)
307
27.1: Classification of Amino Acids. AA’s are classified
according to the location of the amino group.
H H
H2N C C CO2H
H H
H
H2N C CO2H
H
-amino acid
(2-amino carboxylic acid)
-amino acid
(3-amino carboxylic acid)
H H H
H2N C C C CO2H
H H H
-amino acid
(4-amino carboxylic acid)
There are 20 genetically encoded-amino acids found in peptides
and proteins
19 are primary amines, 1 (proline) is a secondary amine
19 are “chiral”, 1 (glycine) is achiral; the natural configuration of
the -carbon is L.
CHO
OH
H
CH2OH
CHO
H
HO
CH2OH
CO2H
H2N
H
CH3
CO2H
H
H2N
R
D-glyceraldehyde
L-glyceraldehyde
CHO
H
HO
H
OH
CH2OH
CHO
OH
H
HO
H
CH2OH
CO2H
H2N
H
H
OH
CH3
CO2H
H
H2N
H3C
H
CH2CH3
D-erythrose
L-erythrose
L-theronine
(2S,3R)
L-isoleucine
(2S,3S)
L-alanine
308
-Amino acids are classified by the properties of their sidechains.
Nonpolar:
COO
COO
COO
–
–
–
NH3
NH3
NH3
Glycine (Gly, G)
(S)-(+)-Alanine (Ala, A)
COO–
COO–
S
COO–
NH3
(S)-(Р)-Leucine (Leu, L)
(S)-(+)-Valine (Val, V)
NH3
NH3
(2S,3S)-(+)-Isoleucine (Ile, I)
(S)-(Р)-Methionine (Met, M)
COO–
COO–
N
H
NH3
H
(S)-(Р)-Proline (Pro, P)
Polar but non-ionizable:
HO
COO–
COO–
N
H
(S)-(Р)-Phenylalanine (Phe, F)
(S)-(Р)-Tryptophan (Trp, W)
COO–
OH
COO–
NH3
NH3
HO
NH3
NH3
(S)-(Р)-Serine (Ser, S)
(2S,3R)-(Р)-Threonine (Thr, T)
(S)-(Р)-Tyrosine (Tyr, Y)
pKa ~ 13
pKa ~ 13
pKa ~ 10.1
HS
COO–
NH3
(R)-(Р)-Cysteine (Cys, C)
pKa ~ 8.2
COO–
H2N
O
NH3
(S)-(Р)-Asparagine (Asn, N)
O
H2N
COO–
NH3
(S)-(+)-Glutamine (Gln, Q)
309
Acidic:
O
COO–
-O
O
NH3
NH3
(S)-(+)-Aspartic Acid (Asp, D)
(S)-(+)-Glutamic Acid (Glu, E)
pKa ~ 3.6
Basic:
H3N
COO–
NH3
COO–
-O
pKa ~ 4.2
N
H
N
H
COO–
NH3
(S)-(+)-Lysine (Lys, K)
(S)-(Р)-Histidine (His, H)
pKa ~ 10.5
pKa ~ 6.0
H
H2N
N
H
N
H
COO–
NH3
(S)-(+)-Arginine (Arg, R)
pKa ~ 12.5
27.2: Stereochemistry of Amino Acids: The natural
configuration of the -carbon is L. D-Amino acids are found in
the cell walls of bacteria. The D-amino acids are not genetically
encoded, but derived from the epimerization of L-isomers
310
27.3: Acid-Base Behavior of Amino Acids. Amino acids exist
as a zwitterion: a dipolar ion having both a formal positive and
formal negative charge (overall charge neutral).
+ R
_
H3N
CO2
H
R
H2N
CO2H
H
pKa ~ 5
pKa ~ 9
Amino acids are amphoteric: they can react as either an acid or a
base. Ammonium ion acts as an acid, the carboxylate as a base.
Isoelectric point (pI): The pH at which the amino acid exists
largely in a neutral, zwitterionic form (influenced by the nature
of the sidechain)
+ R
H3N
CO2H
H
H3O+
pKa1
_
+ R
_
H3N
CO2
H
low pH
Table 27.2 (p. 1115) & 27.2 (p. 1116)
R
HO
H2N
pKa2
CO2
_
H
high pH
311
pKax + pKay
pI =
2
+ CH3
H3N
CO2H
H
low pH
pKa1
(2.3)
+ CH3
H3N
CO2
H
CH3
H2N
CO2
H
pKa2
(9.7)
high pH
CO2H
CO2H
CO2
CO2
CH2
CH2
CH2
CH2
H3N
CO2H
H
pKa1
(1.9)
H3N
CO2
H
pKa3
(3.6)
H3N
CO2
H
pKa2
(9.6)
low pH
H2N
CO2
H
high pH
NH3
NH3
NH3
NH2
(CH2)4
(CH2)4
(CH2)4
(CH2)4
H3N
CO2H
H
low pH
pKa1
(2.2)
H3N
CO2
H
pKa2
(9.0)
H2N
CO2
H
pKa3
(10.5)
H2N
CO2
H
high pH
312
Electrophoresis: separation of polar compounds based on their
mobility through a solid support. The separation is based on
charge (pI) or molecular mass.
+
_
+
_
_
_
_ _
+ +
+
+
313
27.5: Synthesis of Amino Acids:
R-CH2-CO2H
Br
Br2, PBr3
NH2
NH3
R C CO2H
R C CO2H
H
H
Ch. 19.16
Strecker Synthesis: recall reductive amination
NH3
O
NH2
R C CO2H
NH2
NaB(CN)H3
R C CO2H
R C CO2H
H
H
O
NH3
R C H
NH2
NaCN
NH2
R C CN
R C H
H
NH2
H3O+
-orNaOH, H2O
R C CO2H
H
NC:
Amidomalonate Synthesis: recall the malonic acid synthesis
O
O
HN CO2Et
C
H
CO2Et
EtO
Na
RCH2X
HN CO2Et
C
RCH2
CO2Et
H3O
- CO2
H2N H
C
RCH2
CO2H
314
27.5: Reactions of Amino Acids. Amino acids will undergo
reactions characteristic of the amino (amide formation) and
carboxylic acid (ester formation) groups.
H3C
O
H2N
R
H
HOCH2CH3
H3N
H+
R
CO2CH2CH3
H
H3C
O
O
O
CH3
base
CO2
HN
H
R
CO2H
27.6: Some Biochemical Reactions of Amino Acids. Many
enzymes involved in amino acid biosynthesis, metabolism and
catabolism are pyridoxal phosphate (vitamin B6) dependent
(please read)
O
R
H
CO2NH3
+
N
L-amino acid
racemase,
epimerase
N
OH
2-O PO
3
CO2-
R
H
R
NH3
OH
PO
D-amino acid
N
H
pyridoxal
phosphate (PLP)
decarboxylase
R
transaminase
CO2-
R
O
CO2H
H
H
H3N
315
27.7: Peptides. Proteins and peptides are polymers made up of
amino acid units (residues) that are linked together through the
formation of amide bonds (peptide bonds) from the amino
group of one residue and the carboxylate of a second residue
HO
+
H2N
CO2H
H
N
- H2O
H2N
H2N
CO2H
N-terminus
Serine
Alanine
O
CO2H
C-terminus
OH
Ala - Ser
(A - S)
- H2O
HO
N-terminus
H2N
By convention, peptide sequences
are written left to right from the
N-terminus to the C-terminus
C-terminus
H
N
CO2H
O
Ser - Ala
(S - A)
O
H
N
R1
R2
N
H
O
H
N
O
R3
R4
N
H
O
H
N
O
R5
R6
N
H
O
H
N
O
R7
N
H
backbone
316
The amide (peptide) bond has C=N double bond character due
to resonance resulting in a planar geometry
O
H
N
R1
_
R2
N
H
H
N
O
H
N
O
R2
+
N
H
R1
restricts rotations
resistant to hydrolysis
H
N
O
amide bond
The N-H bond of one amide linkage can form a hydrogen bond
with the C=O of another.
O
H N
N-O distance 2.85 - 3.20 Å
R
N H
O
O
N H
N H
O
R
R
H N
optimal N-H-O angle is 180 °
H N
O
O
Disulfide bonds: the thiol groups of cysteine can be oxidized to
form disulfides (Cys-S-S-Cys)
2
HO2C
NH2
H 2O
1/2 O2
NH2
HO2C
SH
H2
S
S
CO2H
NH2
317
R6
N
H
R1
N
H
O
O
HS
O
H
N
R2
H
N
N
H
O
R8
N
H
O
R9
R10
N
H
R9
H
N
N
H
O
R4
R5
N
H
H
N
O
O
H
N
O
R11
N
H
O
O
H
N
O
R12
R13
N
H
H
N
O
S
1/2 O2
SH
H
N
O
O
H
N
R1
H2
N
H
H
N
O
S
O
R2
N
H
O
H
N
O
R4
R5
N
H
H
N
O
Epidermal Growth Factor (EGF): the miracle of mother’s spit
53 amino acid, 3 disulfide linkages
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
1986 Nobel Prize in Medicine:
Stanley Cohen
Rita Levi-Montalcini
318
27.8: Introduction to Peptide Structure Determination.
Protein Structure:
primary (1°) structure: the amino acid sequence
secondary (2°): frequently occurring substructures or folds
tertiary (3°): three-dimensional arrangement of all atoms in a
single polypeptide chain
quaternary (4°): overall organization of non-covalently linked
subunits of a functional protein.
1. Determine the amino acids present and their relative ratios
2. Cleave the peptide or protein into smaller peptide fragments
and determine their sequences
3. Cleave the peptide or protein by another method and
determine their sequences. Align the sequences of the
peptide fragments from the two methods
319
E-A-Y-L-V-C-G-E-R
F-V-N-Q-H-L-F-S-H-L-K
G-C-F-L-P-K
L-G-A
F-V-N-Q-H-L-F
S-H-L-K-E-A-Y
L-V-C-G-E-R-G-C-F
L-P-K-L-G-A
F-V-N-Q-H-L-F
F-V-N-Q-H-L-F-S-H-L-K
S-H-L-K-E-A-Y
E-A-Y-L-V-C-G-E-R
L-V-C-G-E-R-G-C-F
G-C-F-L-P-K
L-P-K-L-G-A
L-G-A
F-V-N-Q-H-L-F-S-H-L-K-E-A-Y-L-V-C-G-E-R-G-C-F-L-P-K-L-G-A
320
27.9: Amino Acid Analysis. automated method to determine the
amino acid content of a peptide or protein
Reaction of primary amines with ninhydrin
O
NH3
R
+
O
O
O
+
N
RCHO
+
CO2
CO2
O
Ninhydrin
peptide
-orprotein
[H]
reduce any
disulfide
bonds
liquid
chromatography
O
O
Enzymatic
digestion
-orH3O+, 
NH3
R
derivatize w/
ninhydrin
Different amino
acids have different
chromatographic
mobilities (retention
times)
CO2
individual
amino acids
Detected w/
UV-vis
1972 Nobel Prize in Chemistry
William Stein
Stanford Moore
321
27.10: Partial Hydrolysis of Peptides. Acidic hydrolysis of
peptides cleave the amide bonds indiscriminately.
Proteases (peptidases): Enzymes that catalyzed the hydrolysis
of the amide bonds of peptides and proteins.
Enzymatic cleavage of peptides and proteins at defined sites:
• trypsin: cleaves at the C-terminal side of basic residues,
Arg, Lys but not His
O
R1
N
H
H3N
O
H
N
R3
N
H
O
H
N
O
CO2
O
R1
trypsin
N
H
H3N
R3
O
H
N
+
O
H3N
CO2
O
O
H2O
H
N
NH3
NH3
• chymotrypsin: cleaves at the C-terminal side of aromatic residues
Phe, Tyr, Trp
R1
O
H3N
N
H
H
N
O
O
R3
N
H
H
N
O
O
CO2
chymotrypsin
H2O
H3N
R1
N
H
H
N
O
O
R3
O
+
H
N
H3N
CO2
O
322
Trypsin and chymotrypsin are endopeptidases
Carboxypeptidase: Cleaves the amide bond of the C-terminal
amino acid (exopeptidase)
27.11: End Group Analysis. The C-terminal AA is identified
by treating with peptide with carboxypeptidase, then analyzing
by liquid chormatography (AA Analysis).
N-labeling: The peptide is first treated with 1-fluoro-2,4-dinitro
benzene (Sanger’s reagent), which selectively reacts with the
N-terminal amino group. The peptide is then hydrolyzed to their
amino acids and the N-terminal amino acid identified as its
N-(2,4-dinitrophenyl) derivative (DNP).
NO2
F
R1
+
H3N
CO2
O
O2N
enzymatic
digestion
-orH3O+, 

H
N
O2N
R1
NO2
N
H
NH2
O
nucleophilic
aromatic
substitution
+
O2N
R1
NO2
N
H
H
N
CO2
O
plus other unlabeled
amino acids
323
27.12: Insulin. (please read) Insulin has two peptide chains
(the A chain has 21 amino acids and the B chain has 30 amino
acids) held together by two disulfide linkages
Pepsin: cleaves at the C-terminal side of Phe, Tyr, Leu; but
not at Val or Ala
Pepsin cleavage
Trypsin cleavage
H3O+ cleavage
324
27.13: The Edman Degradation and Automated Peptide
Sequencing. Chemical method for the sequential cleavage and
identification of the amino acids of a peptide, one at a time starting from
the N-terminus. Reagent: Ph-N=C=S, phenylisothiocyanate (PITC)
Ph
S
C
N
R1
+
H
N
H2N
S
pH 9.0
Ph
CO2
N
H
R1
N
H
O
Ph N
HN
H
N
H+
CO2
O
H+
H+
S H
N
CO2
S
N
Ph
O
HN
OH
R1
R1
H+
N-phenylthiohydantoin:
separated by liquid chromatography
(based of the R group) and detected
by UV-vis
Ph
N
S
+
H2N
CO2
-1 peptide with a new
N-terminal amino acid
(repeat degradation cycle)
O
HN
R1
325
Peptide sequencing by Edman degradation:
• Cycle the pH to control the cleavage of the N-terminal amino
acid by PITC.
• Monitor the appearance of the new N-phenylthiohydantoin for
each cycle.
• Good for peptides up to ~ 25 amino acids long.
• Longer peptides and proteins must be cut into smaller fragments
before Edman sequencing.
Tandem mass spectrometry has largely replaced Edman
degradation for peptide sequencing
27.14: The Strategy for Peptide Synthesis:
Chemical synthesis of peptide:
1. Solution phase synthesis
2. Solid-phase synthesis
326
H
N
H2N
CO2H
- H2O
+
- H2O
H2N
H2N
CO2H
H
N
H2N
CO2H
O
O
Val
Ala
Val - Ala
(V - A)
CO2H
Ala - Val
(A - V)
The need for protecting groups
Pn
+
OH
N
H
peptide
coupling
Pn
OPc
H2N
N
H
Ala - Val
(A - V)
Val
Ala
H
N
H2N
peptide
coupling
(-H2O)
O
OPc
Ala - Val
(A - V)
Pn
Ph
O
Pn
OPc
selectively
remove Pn
O
- H2O
O
O
O
H
N
N
H
H
N
Ph
O
N
H
H
N
O
OPc
Repeat
peptide
synthesis
O
OH
O
Phe (F)
Phe - Ala - Val
(F - A - V)
Orthogonal protecting group strategy: the carboxylate protecting
group must be stable to the reaction conditions for the removal
of the -amino protecting group and ( vice versa)
327
27.15: Amino Group Protection. The -amino group is
protected as a carbamate.
O
NH3
+
O
O
O
Base
RO
RO
NH
Cl
OH
O
O
O
O
O
C6H5
NH
O
O
NH
NH
R
R
O
O
O
tert-butoxycarbonyl
(t-BOC)
benzyloxycarbonyl
(cBz)
fluorenylmethylcarbonyl
(FMOC)
removed with
mild acid
removed with mild acid
or by hydrogenolysis
removed with mild base
(piperidine)
27.16: Carboxyl Group Protection. Protected as a benzyl
ester; removed by hydrogenolysis
O
C6H5
H
N
H2N
O
O
N
H
OH
+
O
H2N
O
C6H5
peptide
coupling
O
C6H5
O
- H2O
O
N
H
H
N
O
mild acid
O
C6H5
O
Ph
O
O
O
C6H5
C6H5
O
peptide
coupling
N
H
OH
O
- H2O
C6H5
O
H
N
O
Ph
O
N
H
H
N
O
O
H2, Pd/C
O
C6H5
O
H3N
Ph
N
H
H
N
O
328
O
O
27.17: Peptide Bond Formation. Amide formation from the
reaction of an amine with a carboxylic acid is slow. Amide bond
formation (peptide coupling) can be accelerated if the carboxylic
acid is activated. Reagent: dicyclohexylcarbodiimide (DCC)
O
O
R
O
C6H11
O
NH
H
R
O
R
+
+
C6H11 N C N C6H11
H
(DCC)
ҐҐ
R'-NH2
C6H11
O
"activated acid"
O
R
R
O C
NH
HN
R'
+ C6H11
O
OH
+
OBn
H2N
O
cBz
peptide
coupling
O
Ala
N
H
R'
N
H
H
N
Ph
cBz
N
H
cBz
OH
O
Phe (F)
H
N
Ph
O
N
H
C6H11
N
H
H
N
OBn
O
H2, Pd/C
OBn
C6H11
DCU
CF3CO2H
H2N
Ph
OBn
O
O
H2N
O
H
N
selectively
remove Nprotecting
group
O
O
N
H
O
Val
DCC
+
Amide
DCC
N
H
O C
+N
N
R'
H
H
C6H11
C6H11
NH
cBz
NH
R
O C
N
C6H11 N C N C6H11
C6H11
O
N
H
H
N
O
OH
O
Phe - Ala - Val
(F - A - V)
329
• In order to practically synthesize peptides and proteins, time
consuming purifications steps must be avoided until the very
end of the synthesis.
• Large excesses of reagents are used to drive reactions forward
and accelerate the rate of reactions.
• How are the excess reagents and by-products from the reaction,
which will interfere with subsequent coupling steps, removed
without a purification step?
27.18: Solid-Phase Peptide Synthesis: The Merrifield Method.
Peptides and proteins up to ~ 100 residues long are synthesized
on a solid, insoluble, polymer support. Purification is conveniently
accomplished after each step by a simple wash and filtration.
330
The solid support (Merrifield resin): polystyrene polymer
Ph
styrene
initiator
Ph
+
polymerization
Ph
Ph
Ph
Ph
Ph
Ph
H3COCH2Cl
ZnCl2
Ph
CH2Cl
Ph
Ph
Ph
Ph
divinylbenzene
(crosslinker, ~1 %)
O
Ph
H
N
_
O
BOC
CF3CO2H
O
R
O
O
R
O
NH
BOC
NH2
R
Solid-phase peptide synthesis
FMOC
O
H2N
DCC
O
Ph
DCC
FMOC
Ph
N
H
O
FMOC
H
N
peptide
coupling
Val
FMOC
OH
N
H
OH
O
Phe (F)
N
H
H
N
O
O
N
H
O
O
O
N
H
O
purify:
wash & filter
O
purify:
wash & filter
N
H
remove Nprotecting
group
N
H
remove Nprotecting
group
HF
remove Nprotecting
group and cleave
from solid-support
purify:
wash & filter
O
H2N
N
H
O
O
purify by liquid
chromatograrphy
Ph
H
N
H2N
or electrophoresis
O
O
331
N
H
OH
O
Ribonuclease A- 124 amino acids, catalyzes the hydrolysis of RNA
Solid-phase synthesis of RNase A:
Synthetic RNase A: 78 % activity
0.4 mg was synthesized
2.9 % overall yield
average yield ~ 97% per coupling step
His-119 A
LYS
GLN
SER
LYS
LYS
LEU
LYS
ASN
ILE
LYS
GLN
GLU
ASP
GLU
HIS
SER
SER
PRO
ALA
ASN
CYS
THR
TYR
ALA
GLY
ALA
THR
MET
SER
ARG
VAL
ASP
VAL
TYR
ASP
PRO
ASN
ASN
SER
ALA
ASP
ASN
ASN
ASN
VAL
ALA
GLN
CYS
ASN
LYS
PRO
VAL
ALA
SER
TYR
LEU
THR
GLN
CYS
SER
ARG
CYS
HIS
TYR
ALA
SER
CYS
THR
PHE
ALA
LYS
TYR
GLU
ALA
ILE
VAL
LYS
THR
ASN
LYS
VAL
VAL
ASN
SER
THR
TYR
ILE
PRO
PHE
SER
GLN
ASP
HIS
CYS
GLY
THR
GLY
LYS
VAL
VAL
GLU
ALA
MET
ARG
GLU
SER
GLN
MET
SER
THR
ALA
HIS
ARG
ALA
MET
CYS
SER
GLN
THR
SER
SER
THR
CYS
PHE
His-12 A
His-12 B
His-119 B
pdb code: 1AFL
R. Bruce Merrifield, Rockefeller University, 1984 Nobel Prize in Chemistry:
“for his development of methodology for chemical synthesis on a solid matrix.”
332
27.19: Secondary Structures of Peptides and Proteins.
-sheet: Two or more extended peptide chain, in which the amide
backbones are associated by hydrogen bonded
anti-parallel
NC
loop
or
turn
H
R
O
N
N O C
R
O
H
R
H
O
O
H
R
O
H
O
O
H
R
H
O
O
R
H
N
N
N
H
R
N
N
N
O
O
N
N
O
R
H
N
N
H
R
N
N
O
CN
H
R
R
O
N
H
R
O
H
R
O
R
H
O
CN
parallel
NC
NC
O
H
R
H
N
O
H
R
R
N
N
N
N
H
R
O
H
R
O
O
H
R
O
R
H
R
H
R
H
O
N
O
N
N
O
R
O
N
N
N
NC
H
R
O
H
crossover
H
N
O
N
N
O
NC
R
H
O
R
333
-helix: 3.6 amino acids per coil, 5.4 Å
C
3.6 AA
5.4 Å
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
N
334
myoglobin
pdb code: 1WLA
Bacteriorhodopsin
pdb code: 1AP9
Parallel -sheets
carbonic anhydrase
pdb code: 1QRM
Anti-parallel
-sheets
of lectin
pdb code: 2LAL
335
27.20: Tertiary Structure of polypeptides and Proteins.
Fibrous. Polypeptides strands that “bundle” to form elongated
fibrous assemblies; insoluble;
Globular. Proteins that fold into a “spherical” conformation .
Hydrophobic effect. Proteins will fold so that hydrophobic amino
acids are on the inside (shielded from water) and hydrophilic
amino acids are on the outside (exposed to water).
Pro • Ile • Lys • Tyr • Leu • Glu • Phe • Ile • Ser • Asp • Ala • Ile • Ile • His •Val • His • Ser • Lys
336
Enzymes: proteins that catalyze biochemical reactions.
• by bringing the reactive atoms together in the optimal geometry
for the reaction.
• lowering the activation energy (G‡) by stabilizing the
transition state and/or high energy intermediate.
• many enzymes use the functional groups of the amino acid
sidechain to carry out the reactions
Proteases (peptidases): catalyzes the hydrolysis of peptide bonds
O
H3N
R
N
H
O
H
N
O
R
R
N
H
O
H
N
O
R
protease
N
H
CO2
H2O
O
H3N
R
N
H
O
H
N
O
R
O
R
O
H
N
+ H N
3
O
R
N
H
CO2
Four classes of proteases:
Serine (trypsin): aspartate-histidine-serine
Aspartyl (HIV protease, renin): two aspartates
Cysteine (papain, caspase): histidine-cysteine
Metallo (Zn2+) (carboxypeptidase, ACE): glutamate
337
Mechanism of carboxylpeptidase, metalloprotease (p. 1151)
Mechanism of a serine protease (trypsin, chymotrypsin):
oxy-anion
hole
NH
NH
HN
NH
O
R1
Ser195
His57
HN
Ser192O
O
N
H
R2
NHR2
R1
O
H
Ser195
His57
N
H
O
N
H
H
N
His57
R1
Ser192O
Ser195
O
H
H
N
H
Asp102 CO2-
N
Asp102 CO2-
O
H
N
acyl-enzyme
intermediate
HN
O
His57
R1
N
H
Asp102 CO2NH
O
H
- R2-NH2
N
H
Asp102 CO2-
HN
O
His57
+
RCO2H
N
N
H
Asp102 CO2-
338
27.21: Coenzymes. Some reactions require additional organic
molecules or metal ions. These are referred to as cofactors or
coenzymes.
S
N
N
+
O
HO
O
P
O
O
OH
P
OH
H
O
HO
P
HO
O
O
O
OH
O
H2N
O
Pyridoxal Phophates
(vitamin B6)
Thiamin Diphosphate
(vitamin B1)
O H
N
N
HN
N
Fe
N
O
CO2CO2-
O
Folic Acid
(vitamin B9)
OH
Heme
NH2
O
OH
O
HN
O
HO
NH
Biotin
(vitamin B7)
CO2H
N
N
O
N
O
O
N
N
H
O
H
OH
Vitamin B12
(cyanocobalamin)
N
O
P O P
O-
OH
S
H
HO
O
-O P
O
O
H
HN
H
N
O
O
N
NH
N
N
HO
NH2
O
O
N
NH2
N
N C N
Co
N
N
H2N
H2N
NH2
O
N
N
NH2
NH2
N
OHO
OH
NH
N
O
Flavin Adenine Diphosphate (FAD)
(Vitamin B2)
27.22: Protein Quaternary Structure. (please read)
339